cDNA Expression Cloning of the IL-1 Receptor, a Member of the Immunoglobulin Superfamily
John E. SimsCaril J. MarchDavid CosmanMichael B. WidmerH. Robson MacDonaldCatherine J. McMahanC. E. GrubinJ WignallJana L. JacksonSusan M. CallDella FriendA AlpertSteven GillisDavid L. UrdalSteven Dower
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Abstract:
Interleukin-1 alpha and -1 beta (IL-1α and IL-1β) are cytokines that participate in the regulation of immune responses, inflammatory reactions, and hematopoiesis. A direct expression strategy was used to clone the receptor for IL-1 from mouse T cells. The product of the cloned complementary DNA binds both IL-1α and IL-1β in a manner indistinguishable from that of the native T cell IL-1 receptor. The extracellular, IL-1 binding portion of the receptor is 319 amino acids in length and is composed of three immunoglobulin-like domains. The cytoplasmic portion of the receptor is 217 amino acids long.Keywords:
Expression cloning
Interleukin-4 receptor
Immunoglobulin superfamily
Cloning (programming)
Immunoglobulin domain
Common gamma chain
Interleukin-4 receptor
Alpha chain
GABBR1
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cDNA clones of the interleukin 1 (IL-1) receptor expressed in a human T-cell clone have been isolated by using a murine IL-1 receptor cDNA as a probe. The human and mouse receptors show a high degree of sequence conservation. Both are integral membrane proteins possessing a single membrane-spanning segment. Similar to the mouse receptor, the human IL-1 receptor contains a large cytoplasmic region and an extracellular, IL-1 binding portion composed of three immunoglobulin-like domains. When transfected into COS cells, the human IL-1 receptor cDNA clone leads to expression of two different affinity classes of receptors, with Ka values indistinguishable from those determined for IL-1 receptors in the original T-cell clone. An IL-1 receptor expressed in human dermal fibroblasts has also been cloned and sequenced and found to be identical to the IL-1 receptor expressed in T cells.
Interleukin-4 receptor
clone (Java method)
GABBR1
Expression cloning
Cell surface receptor
Common gamma chain
Interleukin-1 receptor
Cloning (programming)
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Interleukin-4 receptor
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Interleukin-1 alpha and -1 beta (IL-1α and IL-1β) are cytokines that participate in the regulation of immune responses, inflammatory reactions, and hematopoiesis. A direct expression strategy was used to clone the receptor for IL-1 from mouse T cells. The product of the cloned complementary DNA binds both IL-1α and IL-1β in a manner indistinguishable from that of the native T cell IL-1 receptor. The extracellular, IL-1 binding portion of the receptor is 319 amino acids in length and is composed of three immunoglobulin-like domains. The cytoplasmic portion of the receptor is 217 amino acids long.
Expression cloning
Interleukin-4 receptor
Immunoglobulin superfamily
Cloning (programming)
Immunoglobulin domain
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Citations (896)
Interleukin (IL)-31 is a recently described cytokine, preferentially produced by T helper 2 lymphocytes and associated with skin diseases, such as atopic dermatitis. IL-31 is a member of the four α-helix bundle cytokine family and is related to the IL-6 subgroup. Its heterodimeric membrane receptor is composed of the gp130-like receptor (GPL) subunit associated to the oncostatin M receptor subunit. We identified critical amino acids implicated in the ligand receptor interaction by computational analysis combined with site-directed mutagenesis. Six IL-31 residues selected for their putative involvement in cytokine receptor contact sites were alanine-substituted, and the corresponding proteins were expressed in mammalian and bacterial systems. Biochemical, membrane binding, cell signaling, and cell proliferation analyses showed that mutation E44A, E106A, or H110A abolished IL-31 binding to GPL and the subsequent signaling events. A second ligand receptor-binding site involved Lys134, with alanine substitution leading to a protein that still binds GPL, but is unable to recruit the second receptor subunit and the subsequent signaling pathways. The results indicate that IL-31 recognizes its receptor complex through two different binding sites, and we propose a three-dimensional model for IL-31. Interleukin (IL)-31 is a recently described cytokine, preferentially produced by T helper 2 lymphocytes and associated with skin diseases, such as atopic dermatitis. IL-31 is a member of the four α-helix bundle cytokine family and is related to the IL-6 subgroup. Its heterodimeric membrane receptor is composed of the gp130-like receptor (GPL) subunit associated to the oncostatin M receptor subunit. We identified critical amino acids implicated in the ligand receptor interaction by computational analysis combined with site-directed mutagenesis. Six IL-31 residues selected for their putative involvement in cytokine receptor contact sites were alanine-substituted, and the corresponding proteins were expressed in mammalian and bacterial systems. Biochemical, membrane binding, cell signaling, and cell proliferation analyses showed that mutation E44A, E106A, or H110A abolished IL-31 binding to GPL and the subsequent signaling events. A second ligand receptor-binding site involved Lys134, with alanine substitution leading to a protein that still binds GPL, but is unable to recruit the second receptor subunit and the subsequent signaling pathways. The results indicate that IL-31 recognizes its receptor complex through two different binding sites, and we propose a three-dimensional model for IL-31.
Glycoprotein 130
Interleukin-4 receptor
Cytokine receptor
Oncostatin M
Interleukin-13 receptor
Interleukin-6 receptor
Common gamma chain
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The α-subunit of interleukin-6 (IL-6) receptor is a member of the hematopoietin receptor family. The alignment of its amino acid sequence with those of other members of this family (human somatotropin receptor/murine IL-3 receptor β and human IL-2 receptor β) has suggested that amino acids included in two SSFY repeats found in each of its hematopoietin receptor domains, contribute to the binding of the ligand. The involvement of these amino acids in IL-6 binding and signal transduction was studied by site-directed mutagenesis and molecular modelling. We present a computer-derived three-dimensional model of the IL-6/IL-6 receptor complex based on the structure of the human somatotropin/human somatotropin receptor complex. This model allowed the location of distinct regions important for IL-6 and gp130 binding. We show that some of the residues included in the SSFY repeats located in our IL-6 receptor model in the loops between β-strands E and F of domain-I and B' and C', of domain-II, participate in the formation of a major IL-6-binding site. These residues are necessary for IL-6 and gp130 binding and for signal transduction. Using our IL-6 receptor mutants we mapped the epitopes of four anti-(IL-6 receptor) neutralising monoclonal antibodies to these residues. Our results demonstrate that a generic hematopoietin receptor family structural module can be used for the study of both α and β receptor subunits belonging to this family.
Glycoprotein 130
Interleukin-4 receptor
GABBR1
Interleukin-13 receptor
Interleukin-6 receptor
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Interleukin-2 (IL-2), a multifunctional cytokine, plays a major role on immune reactions through combining with its receptor on surface of T cells. Interleukin-2 receptor, an important immune signal transmissible molecule, consists of three subunits, such as α, β and γ. The α subunit is a low affinity receptor, the β and γ subunits together form an intermediate affinity receptor, which converts to a high affinity form with the co-expression of the α subunit. All of that can be expressed on surface of T cells. Activation of T cells can be induced by interleukin-2 receptor through signalling pathway of STAT, MAPK, PI3K, and plays key role on antiviral and antibacterial immune reactions.
Interleukin-4 receptor
Immune receptor
Cytokine receptor
Interleukin-6 receptor
Interleukin-1 receptor
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The α‐subunit of interleukin‐6 (IL‐6) receptor is a member of the hematopoietin receptor family. The alignment of its amino acid sequence with those of other members of this family (human somatotropin receptor/murine IL‐3 receptor β and human IL‐2 receptor β) has suggested that amino acids included in two SSFY repeats found in each of its hematopoietin receptor domains, contribute to the binding of the ligand. The involvement of these amino acids in IL‐6 binding and signal transduction was studied by site‐directed mutagenesis and molecular modelling. We present a computer‐derived three‐dimensional model of the IL‐6/IL‐6 receptor complex based on the structure of the human somatotropin/human somatotropin receptor complex. This model allowed the location of distinct regions important for IL‐6 and gp130 binding. We show that some of the residues included in the SSFY repeats located in our IL‐6 receptor model in the loops between β‐strands E and F of domain‐I and B' and C', of domain‐II, participate in the formation of a major IL‐6‐binding site. These residues are necessary for IL‐6 and gp130 binding and for signal transduction. Using our IL‐6 receptor mutants we mapped the epitopes of four anti‐(IL‐6 receptor) neutralising monoclonal antibodies to these residues. Our results demonstrate that a generic hematopoietin receptor family structural module can be used for the study of both α and β receptor subunits belonging to this family.
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Interleukin-4 receptor
Interleukin 3
Cytokine receptor
Gamma subunit
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Interleukin-1 receptor
Interleukin-4 receptor
Expression cloning
Cloning (programming)
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